Antigenic and genotypic characterization of rabies virus isolated from bats (Mammalia: Chiroptera) from municipalities in São Paulo State, Southeastern Brazil

Bats have aroused growing attention in the public health sphere because they are considered the main reservoir of rabies virus (RABV) in the Americas, in places where canine rabies is under control. Antigenic and genetic studies of RABV isolates have been used to describe the epidemiological profile of rabies and to identify possible hosts/reservoirs for different epidemiological cycles. This study describes the antigenic and genotypic characterization of 19 RABV isolates from central nervous system samples of non-hematophagous bats (Mammalia: Chiroptera). These bats were diagnosed as RABV positive by direct fluorescent antibody and mouse inoculation tests. Antigenic characterization using a panel of eight monoclonal antibodies revealed that 7 of 19 RABV isolates from these bats belonged to variant 3, for which the hematophagous bat species Desmodus rotundus is the main reservoir, and 1 of 19 RABV isolates from an insectivorous bat belonged to variant 4, which is characteristic of these bats. The remaining 11 RABV samples were divided into six non-compatible profiles. The isolates were subjected to reverse transcription polymerase chain reaction for the N gene and partially sequenced. Genetic characterization of these isolates was performed by grouping the sequences obtained with known RABV lineages. The sequences were grouped in clusters by the phylogenetic inference neighbor-joining method, together with another 89 homologous sequences obtained from GenBank. This analysis grouped the isolates into four known lineages: Nyctinomops Brazil, Myotis Brazil, Eptesicus Brazil and D. rotundus Brazil, as well as another cluster that may define a RABV lineage not yet characterized, here named Myotis Brazil II, for which bats of the genus Myotis apparently act as reservoirs. This assumption of a new lineage is also based on the observation of amino acid substitutions, with an average intraspecific identity of 99.8%, varying from 99.6 to 100.0% for nucleotides and 100.0% for amino acids.